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Aim To investigate the inhibition effect of 2-dodecyl-6-methoxycyclohexa-2, 5-diene-l, 4-dione ( DMDD) on renal tubular epithelial cell HK-2 endo¬plasmic reticulum stress and inflammatory responses induced by high glucose. Methods HK-2 cells were cultured in vitro and divided into normal group, high glucose group, endoplasmic reticulum stress inhibitor 4-PBA group (5 mmoL • L ) , DMDD high, medium and low dose groups (8,4,2 μmol • L
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Objective@#Doubled blepharoplasty, which has a various types, is one of the commonest plastic surgery procedures. Incision method has a long-standing result but linear scars can be seen when eyes are closed. So it is necessary to improve this surgerical procedure. The aim of this report is to propose a novel palpebral margin incision technique which avoids scars in the upper eyelid.@*Methods@#15 patients undertook blepharoplasty with palpebral margin incision technique. All the patients in this study were healthy with no other serious diseases. The double-eyelid crease was formed after the operation.The aesthetic results were evaluated based on their satisfaction.@*Results@#All of the patients were satisfied with aesthetic results, and no significant complications occurred. And No obvious regression of the double fold took place.@*Conclusions@#This new method provides an effective way to achieve eyelid reconstruction without obvious scar in the upper eyelid which is of very important clinical significance and makes cosmetic surgery process take a step forward.
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High-resolution melting (HRM) analysis is a versatile method for variant scanning and genotyping. It involves amplification of the target in the presence of a saturation dye by the polymerase chain reaction (PCR). HRM analysis can be performed in one closed-tube, which does not require additional post-PCR separations and greatly reduces the possibility of contamination. HRM is faster, simpler, and less expensive than alternative approaches requiring labeled probes. Considering the many advantages of HRM analysis, many researchers have tried to apply this method to forensic research. This paper intends to summarize the principle, technical characteristics, limitations and application of HRM analysis in forensic science.
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Objective To investigate if there were connections between sporadic Parkinson's disease (PD) and three single nucleotide polymorphisms (SNPs) in transmembrane protein 175 (TMEM175 rs34311866), methylcrotonoyl-coenzyme A carboxylase 1 ( MCCC1 rs12637471 ) and alpha-synuclein (SNCA rs356182) in Northern Chinese Han population , and provide basic data for PD genetic research. Methods The research recruited 310 sporadic PD patients in northern Chinese Han population from the Department of Neurology, the First Hospital of China Medical University between 2008 and 2012, and 339 controls without nervous system manifestations from other departments of the First Hospital of China Medical University during the same period.We applied cleaved amplification polymorphism sequence-tagged sites polymerase chain reaction-restriction fragment length polymorphism method to detect the genotype distributions of the SNPs in the northern Chinese Han population , and calculated relevance with PD of the SNPs by chi-square test.Results According to the data, the allele A of SNCA rs356182 had positive effects on the onset of PD in northern Chinese Han population compared with controls (patient group A%=20.97%(130/620), control group A% =29.20%(198/678), χ2=11.632, P=0.001); allele G of MCCC1 rs12637471 (χ2=0.009, P=0.926) and allele C of TMEM175 rs34311866 (χ2=1.369, P=0.242) showed no significant differences between PD and control groups.Conclusion SNCA rs356182 was related with PD, and TMEM175 rs34311866 (M311Y) as well as MCCC1 rs12637471 showed no correlation with PD in the northern Chinese Han population.
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Objective To explore the potential relationship between 6 single nucleotide polymorphisms (SNPs) of mitochondrial DNA (mtDNA) coding region and Parkinson's disease (PD) in northern Chinese population.Methods Samples from 353 patients with PD and 389 control participants,collected in our hospital from April 2010 to November 2015,were genotyped by multiplex polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) techniques.Six SNPs (T4216C within the ND1 gene,A4917G within the ND2 gene,A11084G within the ND4 gene,A12308G within the tRNA gene,A13966G within the ND5 gene,and G15928A within the tRNA gene) in mtDNA were amplified in multiplex PCR systems and subsequently genotyped by digestion with endonucleases.Results An efficient,rapid,and economical way of multiplex genotype SNPs was successfully established.No significant differences were observed in the 6 loci and corresponding haplotypes between PD and control cohorts (P>0.05).After stratification by gender,however,frequencies of allele 11084G and the haplotype T-A-G-A-A-G in female group with PD were all higher than those in control ones,with significant differences (P<0.05).Conclusions The 6 SNPs may not be susceptible factors in the northern Chinese population.The A11084G missense mutation might be related to PD in female patients.
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Objective To investigate personal identification of mixed seminal stain of two individuals, we combined the detection of genotyping autosomal, Y and X STR and sequencing mtDNA hypervariable Ⅰ (HV Ⅰ ) region. Methods We analyzed autosomal, Y and X STR with commercial kit and separating and sequencing HVⅠfragments of mixed seminal stain from two males by SSCP electrophoresis. Results Four genetic markers of the high amount sample can be obtained when mixed ratio is more than 1:10. When the proportion of two samples is close, the suspect could be excluded or, to some extent, identified by comparing with our results. Conclusion The combined detection of four genetic marker systems can, to some degree, solve the personal identification from mixed seminal stain of two individuals.
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HTR1A is a kind of receptor subtypes to express the most 5-HT in the brain of mammalian. Genetic variation in the gene through influencing the expression of receptors or changing the activity of the ligand binding to increase or decrease the function of 5-HT, so it played a key role in the pathogenesis of depression and other mental system diseases. The article reviewed the association between the genetic mutations of HTR1A and the corresponding mental system disease. Research the relationship of HTR1A gene and mood changes such as depression, panic, impulsive personality and its significance in forensic judicial authentication. To explore the causes of criminal mode from the perspective of gene - psychology - social model.
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Objective In this study,a multiplex PCR amplification system was constructed based on fluorescent labeling PCR and LDR,to provide a new strategy for analyzing severely degraded DNA.Methods Eight SNP loci (rs10802248,rs10516197,rs10488372,rs2278945,rs4757318,rs4887255,rs4889002,and rs9304473) were selected.Their LDR probes and PCR primers of linked products were designed and synthesized.Ligase detection reaction,PCR amplification,and capillary gel electrophoresis (CEG) were performed to establish the multiplex LDR-PCR amplification system.Results The genotypes of these 8 loci were obtained simultaneously by the fluorescence-labeled multiplex LDR-PCR amplification method.The loci profiles obtained by fluorescence-labeled multiplex LDR-PCR amplification were in accordance with those obtained by direct sequencing of the polymorphic regions in samples from all individuals.By fluorescence-labeled multiplex LDR-PCR amplification,the 8 SNP loci were efficiently amplified from the severely degraded FFPET DNA.Conclusion Eight SNP loci results could be obtained simultaneously by using the multiplex LDR-PCR amplification system,which is a simple,efficient,and practical SNP genotyping method with accurate and reliable results for highly degraded samples.
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Objective To investigate SNP and distribution of haplotypes in differentially m ethylated region (DMR) upstream of H19 gene in Chinese Korean nationality in order to provide basic data for forensic application and population genetics research. Methods One hundred and one blood sam ples from unrelat-ed Chinese Korean individuals and 14 blood sam ples from 5 Chinese Korean intergenerational fam ilies which known genetic relationship were collected. The SNP in DMRupstream of H19 gene were investi-gated by PCR-cycle sequencing and McrB Cdigestion followed by PCR . The haplotypes detected by parentally im printed allele (PIA) m ethod and relevant genetic param eters were calculated. Results Thirteen SNPs (rs10840167, rs2525883, rs12417375, rs4930101, rs2525882, rs2735970, rs2735971, rs11042170, rs2735972, rs10732516, rs2071094, rs2107425, and rs4930098) and five haplotypes were detected in 1 174 bp target product in DMRupstream of H19 gene, with 9 SNPs having high discrim ination power as good genetic markers. The average gene diversity (GD) of haplotypes was 0.714. The maternal haplotype was confirm ed correctly by PIAm ethod from McrB C-digested products of genom ic DNA. Conclusion High polym orphism sexist in DMRupstream of H19 gene in Chinese Korean nationality. And determ ination of the maternal haplotype could furtherm ore enhance the forensic identification efficiency of im printed gene.
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OBJECTIVE@#To investigate the genetic polymorphism in the 5' and 3' region of TPH2 gene of Northern Chinese Han population and to explore its application value in forensic medicine.@*METHODS@#The sequence variants and the genetic polymorphisms of 6 SNP loci (rs4570625, rs11178997, rs11178998, rs41317118, rs17110747 and rs41317114) within a 905 bp 5' flanking region and a 1,104bp 3' flanking region of TPH2 gene were analyzed by DNA sequencing in a total of 244 unrelated healthy individuals in Northern Chinese Han population. The statistical analysis was carried out by Haploview v4.2 software.@*RESULTS@#The genotypic distributions of the 6 SNP loci in the TPH2 gene were in accordance with Hardy-Weinberg equilibrium. One C/T variant in 92922 site was found. There was a high linkage disequilibrium among the 3 SNP loci (rs4570625, rs11178997 and rs11178998) in the 5' region and the 3 SNP loci (rs41317118, rs17110747 and rs41317114) in the 3' region of TPH2 gene, respectively. The parameters of population genetics of 6 SNP loci were obtained.@*CONCLUSION@#There are great polymorphisms in the 5' and 3' region of TPH2 gene in Northern Chinese Han population, which could be used as genetic indexes for association analysis of the related diseases, as well as for forensic individual identification and paternity testing.
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Humans , 3' Untranslated Regions , 5' Untranslated Regions , Asian People/genetics , China/ethnology , Forensic Genetics , Gene Frequency , Genetics, Population , Genotype , Linkage Disequilibrium/genetics , Microsatellite Repeats , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Tryptophan Hydroxylase/geneticsABSTRACT
OBJECTIVE@#To reveal the genetic polymorphisms of four SNP loci (rs77434921, rs2076907, rs6283, rs1800762) in D5 gene of dopamine receptor (DRD5) in Northern Chinese Han population.@*METHODS@#Four SNP loci of the DRD5 gene of 206 unrelated individuals in Northern Chinese Han population were separately amplified and sequenced by PCR technique and statistically analyzed by Haploview v4.1 software.@*RESULTS@#In Northern Chinese Han population, the genotype frequency distribution of rs77434921, rs2076907, rs6283 and rs1800762 loci in the DRD5 gene were all in accordance with Hardy-Weinberg equilibrium. DP value was 0.145, 0.532, 0.602 and 0.159, while PE value was 0.004, 0.079, 0.196 and 0.007. A linkage disequilibrium among these four SNP loci was also demonstrated, which might infer five haplotypes.@*CONCLUSION@#rs2076907 and rs6283 loci of DRD5 gene in the Northern Chinese Han population have high genetic polymorphisms, which can be useful for forensic identification of individuals.
Subject(s)
Humans , Asian People/genetics , China/ethnology , DNA Primers/genetics , Forensic Genetics , Gene Frequency , Genetic Markers , Genotype , Haplotypes , Linkage Disequilibrium , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Receptors, Dopamine D5/geneticsABSTRACT
OBJECTIVE@#To investigate the single nucleotide polymorphisms (SNP) of -855 G/C and -1140 G/A in promoter regions of GRIN1 gene and find their genetic correlation to paranoid schizophrenia as well as their applicable values in forensic medicine.@*METHODS@#The genetic polymorphisms of -855 G/C and -1140 G/A at the 5' end of GRIN1 gene were detected by PCR restriction fragment length polymorphism and PAGE in 183 healthy unrelated individuals of northern Chinese Han population and 172 patients of paranoid schizophrenia, respectively. The chi2 test was used to identify Hardy-Weinberg equilibrium of the genotype distribution. The differences of genotypes and allelic frequency distributions were compared between the two groups.@*RESULTS@#Distributions of the genotypic frequencies satisfied Hardy-Weinberg equilibrium in both groups. The difference of genotypes was statistically significant between female patient group and female control group in -855 G/C distribution (P < 0.05). The differences of genotypes and allelic frequencies were statistically significant not only between the patient group and the control group but also between female patient group and female control group in -1140 G/A distribution (P < 0.05).@*CONCLUSION@#The SNP of -1140 G/A in promoter regions of GRIN1 gene might positively correlate to paranoid schizophrenia. The genetic factor of schizophrenia is involved in gender tendency. And it could be useful in forensic identification of schizophrenia.
Subject(s)
Female , Humans , Male , Alleles , Asian People/genetics , Base Sequence , Gene Frequency , Genetic Predisposition to Disease/genetics , Genotype , Nerve Tissue Proteins/genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Receptors, N-Methyl-D-Aspartate/genetics , Schizophrenia, Paranoid/genetics , Sequence Analysis, DNAABSTRACT
OBJECTIVE@#To explore the feasibility of improving the sensitivity of DNA detection by increasing the PCR cycle index and decreasing the volume of amplifying system.@*METHODS@#The DNA of semen were collected from 10 healthy irrelevant volunteers, and were quantified to 50, 40, 30, 25, 20, 15, 10 pg/microL, separately. All samples were then amplified in 10, 5, 3 microL volume and at 28, 30, 32, 34, 36 cycles, respectively. 3130 genetic analyzer was used to detect 15 autosomal STR loci.@*RESULTS@#Under the situation of 28 cycles and 3 microL volume, samples which achieved > 40 pg/microL could be correctly typed. Under the situation of 10, 5, 3 microL volume, samples which achieved > 20 pg/microL could be correctly typed at 34 cycles. When increasing the index to 36 cycles, they could not be correctly typed because of the non-specific band.@*CONCLUSION@#DNA detecting sensitivity can be improved to a certain extent by increasing the cycle index and decreasing the volume of amplifying system.
Subject(s)
Humans , Male , DNA/genetics , DNA Fingerprinting/methods , Feasibility Studies , Forensic Genetics/methods , Limit of Detection , Polymerase Chain Reaction/methods , Semen/chemistry , Sensitivity and Specificity , Tandem Repeat SequencesABSTRACT
To investigate the association of five SNPs (rs823083, rs708723, rs4951261, rs823076 and rs16856110) at the PARK16 locus with Parkinson's disease (PD), and to potentiate its forensic application. The genomic DNAs of 215 PD patients and 212 matched controls from the northern Han Chinese population were amplified in two independent PCR systems and subsequently genotyped by digestion with the three endonucleases (Hinf I, Nco I and Msp I ). The genetic parameters and association studies were carried out with SPSS 13.0, Haploview version 4.2 and PLINK 1.07 softwares. We detected accurately all genotypes in the five SNPs with multiplex PCR-RFLP and mismatched multiplex PCR-RFLP techniques. The genotypes of four SNPs, except for rs823083, were in Hardy-Weinberg equilibrium. The four SNPs, rs16856110, rs4951261, rs708723 and rs823076, which were in linkage equilibrium, should not be associated with PD (P-values ranging from 0.077 to 0.544). The SNPs investigated at the PARK16 locus were not found to be involved in PD-associated blocks in the northern Han Chinese population. The allele distributions of rs708723, rs4951261, rs823076 and rs16856110 in the northern Han Chinese population can be highly polymorphic, which can be applied to genetic analysis and forensic practices.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Asian People/genetics , Case-Control Studies , Forensic Genetics , Gene Frequency , Genetic Association Studies , Genetic Loci , Genetic Predisposition to Disease , Genotype , Parkinson Disease/genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single NucleotideABSTRACT
OBJECTIVE@#To investigate SNP and distribution of haplotypes in differentially methylated region (DMR) upstream of H19 gene in Chinese Korean nationality in order to provide basic data for forensic application and population genetics research.@*METHODS@#One hundred and one blood samples from unrelated Chinese Korean individuals and 14 blood samples from 5 Chinese Korean intergenerational families which known genetic relationship were collected. The SNP in DMR upstream of H19 gene were investigated by PCR-cycle sequencing and McrBC digestion followed by PCR. The haplotypes detected by parentally imprinted allele (PIA) method and relevant genetic parameters were calculated.@*RESULTS@#Thirteen SNPs (rs10840167, rs2525883, rs12417375, rs4930101, rs2525882, rs2735970, rs2735971, rs11042170, rs2735972, rs10732516, rs2071094, rs2107425, and rs4930098) and five haplotypes were detected in 1 174 bp target product in DMR upstream of H19 gene, with 9 SNPs having high discrimination power as good genetic markers. The average gene diversity (GD) of haplotypes was 0.714. The maternal haplotype was confirmed correctly by PIA method from McrBC-digested products of genomic DNA.@*CONCLUSION@#High polymorphisms exist in DMR upstream of H19 gene in Chinese Korean nationality. And determination of the maternal haplotype could furthermore enhance the forensic identification efficiency of imprinted gene.
Subject(s)
Humans , Asian People/genetics , China , DNA/genetics , DNA Methylation , DNA Primers , Forensic Genetics/methods , Gene Frequency , Genotype , Haplotypes , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , RNA, Long Noncoding/genetics , Republic of Korea/ethnology , Sequence Analysis, DNAABSTRACT
<p><b>BACKGROUND</b>Parkinson's disease (PD) is an autosomally inherited neurodegenerative disease in elderly people. The etiology of PD has long been thought to be associated with both genetic and environmental factors. To explore potential genetic risk factors for PD in the northern Han Chinese population, we investigated three single nucleotide polymorphisms (SNPs) (rs4538475, rs11107 and rs12564040) in the BST1, PARK15 and PARK9 genes.</p><p><b>METHODS</b>Genomic DNA from 215 PD patients and 212 matched controls was amplified in two independent PCR systems and subsequently genotyped by digestion with the endonuclease PstI. Genetic parameter and association studies were carried out with SPSS 13.0 and PLINK 1.07 software.</p><p><b>RESULTS</b>We could accurately detect all genotypes in the three loci with the PCR-RFLP or mismatched PCR-RFLP techniques. The observed heterozygosities of the rs4538475 and rs11107 loci in PD and control groups ranged from 0.460 - 0.481 and 0.410 - 0.441, in BST1, PARK15 respectively, while we detected no heterozygosity at the rs12564040 locus in PARK9. The similar distributions of genotypic frequency between both groups suggest that the three SNPs investigated in this study are unlikely to play roles as common risk factors or pathogenic mutations for PD in northern Han Chinese.</p><p><b>CONCLUSION</b>The SNPs investigated in the BST1, PARK15 and PARK9 genes associated with PD susceptibility are not associated with PD in the northern Han Chinese population.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , ADP-ribosyl Cyclase , Genetics , Antigens, CD , Genetics , Asian People , Genetics , F-Box Proteins , Genetics , GPI-Linked Proteins , Genetics , Genetic Predisposition to Disease , Genetics , Parkinson Disease , Genetics , Parkinsonian Disorders , Genetics , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polymorphism, Single Nucleotide , GeneticsABSTRACT
OBJECTIVE@#To investigate distribution specificity of human fucosyltransferase 5 (FUT5) as well as its expression and localization in spermatids.@*METHODS@#Human semen, vaginal swab, saliva and venous blood from healthy individuals were collected. The spermatids were isolated and the spermatid membrane protein was then extracted. Expression levels of FUT5 from human spermatid membrane, seminal plasma, vaginal fluid, saliva and serum were detected by immunoblotting technique. The expression and localization of FUT5 in spermatids were analyzed by immunofluorescent method.@*RESULTS@#Immunoblotting technique showed that FUT5 was expressed on spermatid membranes and in serum, but not in seminal plasma, vaginal fluid and saliva. The expressed FUT5 on spermatids was mostly localized on head of spermatids by fluorescent microscopy, suggesting that there was certain amount of FUT5 on human spermatid membrane, and the spermatids might be isolated from mixed stains with vaginal fluid by antigen-antibody reaction.@*CONCLUSION@#Human FUT5 shows a characteristic distribution specificity, and this feature may be used for identification of mixed stain involved in criminal sexual offence in future forensic practice.
Subject(s)
Female , Humans , Male , Cell Membrane/metabolism , Fluorescent Antibody Technique/methods , Forensic Genetics/methods , Fucosyltransferases/metabolism , Immunoblotting , Saliva/metabolism , Semen/metabolism , Spermatids/metabolism , Vagina/metabolismABSTRACT
OBJECTIVE@#To investigate the polymorphisms of rs4906902 and rs8179184 loci in the promoter of the gamma-aminobutyric acid(GABA) receptor A, beta3 subunit gene (GABRB3), and their relevance with schizophrenia.@*METHODS@#PCR and DNA sequencing were used to detect the polymorphisms of rs4906902 and rs8179184 loci in 210 healthy individuals (control group) and 206 schizophrenic patients (case group) of the Han population in northern China. The chi2 test was used to identify Hardy-Weinberg equilibrium of the genotype distribution in the control group followed by comparing differences in genotype and haplotype frequency distributions between two groups.@*RESULTS@#Distributions of the genotype frequencies fit the law of Hardy-Weinberg equilibrium in the control group. rs4906902 and rs8179184 loci were in linkage disequilibrium and showed two haplotypes which were T-G and C-A. The differences of genotypic frequencies and haplotype frequencies were statistically significant between the two groups (P < 0.05). The frequency of haplotype C-A in the case group was significantly higher than in the control group. Genotypic and haplotype frequencies in the maternal line and paternal line were statistically significant in the case group (P < 0.05).@*CONCLUSION@#The haplotype of C-A in rs4906902 and rs8179184 loci in the promoter of GABRB3 gene may be maternally inherited and positively associated with schizophrenia and may be a useful tool in the forensic identification of schizophrenia.
Subject(s)
Female , Humans , Male , Alleles , Asian People/genetics , China/epidemiology , Gene Frequency , Genetic Predisposition to Disease , Genotype , Haplotypes , Inheritance Patterns , Linkage Disequilibrium , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Promoter Regions, Genetic , Receptors, GABA-A/genetics , Schizophrenia/genetics , Sequence Analysis, DNAABSTRACT
Catechol-O-methyltransferase (COMT) gene encodes catechol-O-methyltransferase, the variant of this gene may affect the expression and metabolic activity of COMT. As the result of the changes of the effective concentration of the catecholamine neurotransmitter in the central nervous system, central nervous system dysfunctions associated with schizophrenia. This review summarizes genetic polymorphism and diversity of COMT gene. It also elaborates the relation between SNP and haplotype of COMT gene and three aspects, which including schizophrenia, attacking and violent tendency, and the frontal cognitive function of the schizophreniac. The correlativity study between genetic variation of the COMT gene and schizophrenia in patients with attacking and violent tendency may be helpful for the assessment of forensic psychiatry.
Subject(s)
Humans , Aggression/psychology , Brain/pathology , Catechol O-Methyltransferase/genetics , Cognition/physiology , Dopamine/metabolism , Forensic Genetics , Gene Expression , Genetic Predisposition to Disease , Genetic Variation , Genotype , Haplotypes , Polymerase Chain Reaction , Polymorphism, Genetic , Prefrontal Cortex/pathology , Promoter Regions, Genetic , Schizophrenia/genetics , Violence/psychologyABSTRACT
OBJECTIVE@#To investigate the population genetic data of 3 SNP loci (rs25533, rs34388196 and rs1042173) of 5-hydroxytryptamine transporter (5-HTT) gene and the association with paranoid schizophrenia.@*METHODS@#Three SNP loci of 5-HTT gene were examined in 132 paranoid schizophrenia patients and 150 unrelated healthy individuals of Northern Chinese Han population by PCR-RFLP technique. The Hardy-Weinberg equilibrium test was performed using the chi-square test and the data of haplotype frequency and population genetics parameters were statistically analyzed.@*RESULTS@#Among these three SNP loci, four haplotypes were obtained. There were no statistically significant differences between the patient group and the control group (P > 0.05). The DP values of the 3 SNP loci were 0.276, 0.502 and 0.502. The PIC of them were 0.151, 0.281 and 0.281. The PE of them were 0.014, 0.072 and 0.072.@*CONCLUSION@#The three SNP loci and four haplotypes of 5-HTT gene have no association with paranoid schizophrenia, while the polymorphism still have high potential application in forensic practice.